Using 16 human gingival fibroblast cell lines from patients with periodontitis, Dilantin hyperplasia, and nonpathological gingiva, a microscopic assay was developed to quantitate the cells' ability to lyse collagen substrates. The method employs tissue culture chambers with one cover slip partially coated with a thin layer of undenatured fibrillar bovine codlagen. The assay measures the relative numbers and sizes of holes in the collagen within defined regions of the cover slips effected by the phagocytotic and collagenolytic performance (PCP) of the population of fibroblasts growing on the cover slip for 5 days. The effect on the PCP index by serum, heparin, prostaglandins, and endotoxin was evaluated.
Get full access to this article
View all access options for this article.
References
1.
Rose, G.G. and Robertson, P.B.: Collagenolysis by Human Gingival Fibroblast Cell Lines, J Dent Res56:416-424, 1977.
2.
Yajima, T. and Rose, G.G.: Phagocytosis of Collagen by Human Gingival Fibroblasts In Vitro, J Dent Res56:1271-1277, 1977.
3.
YAJIMA, T. and ROSE, G. G.: in preparation.
4.
Eley, B.M. and Harrison, J.D.: Intracellular Collagen Fibrils in the Periodontal Ligament of Man, J Periodont Res10:168-170, 1975.
5.
Beertsen, W. ; Everts, J.; and Van DenHoof, A.: Fine Structure of Fibroblasts in the Periodontal Ligament of the Rat Incisor and Their Possible Role in Tooth Eruption, Archs Oral Biol19:1087-1098, 1974.
6.
Garant, P.R. : An Electron Microscopic Study of the Periodontal Tissues of Germfree Rats and Rats Monoinfected with Actinomyces naeslundii , J Periodont Res15:36-72, 1976.
7.
Listgarten, M.A.: Intracellular Collagen Fibrils in the Periodontal Ligament of the Mouse, Rat, Hamster, Guinea Pig, and Rabbit, J. Periodont Res8:335-342, 1973.
8.
Ten Cate, A.R. and Deporter, D.A.: The Role of the Fibroblast in Collagen Turnover in the Functioning Periodontal Ligament of the Mouse, Archs Oral Biol19: 339-340,1974.
9.
Ten Cate, A.R. ; Deporter, D.A.; and Freeman, E.: The Role of Fibroblasts in the Remodeling of Periodontal Ligament During Physiologic Tooth Movement, Am. J Orthodont69:155-168, 1976.
10.
Soames, J.V. and Davies, R.M.: Intracellular Collagen Fibrils in Early Gingivitis in the Beagle Dog, J Periodont Res12 :378-386, 1977.
11.
Golub, L.M. ; Garant, P.R.; and Rama-Murthy , N.S.: Inflammatory Changes in Gingival Collagen in the Alloxan-Diabetic Rat, J Periodont Res12:402- 418, 1977.
Rose, G.G.; Kumegawa, M.; Nikai H.; Bracho, H.; and Cattoni, M.: The Dual-rotary Circumfusion System for Mark II Culture Chambers. I. Design, Control, and Monitoring of the System and the Cultures, Microvascular Res2:24-60, 1970.
14.
Fullmer, H.M. ; Taylor, R.E. and Guthrie, R.W.: Human gingival collagenase: purification, molecular weight, and inhibitor studies, J Dent Res51:349-355, 1972.
15.
Birkedal-Hansen, H.; Cobb, C.M.; Taylor, R.E. and Fullmer, H.M.: Serum Inhibition of Gingival Collagenase , J Oral Path3:284-290, 1974.
16.
Taylor, A.C. : Collagenolysis in Cultured Tissue: II. Role of Mast Cells, J Dent Res50:1301-1306, 1971.
17.
Sakamoto, S. ; Goldhaber, P. and Glimcher, M.J.: Mouse Bone Collagenase: The Effect of Heparin on the Amount of Enzyme Released in Tissue Culture and on the Activity of the Enzyme, Calc Tiss Res12: 247-258, 1973.
18.
Goodson, J.M. ; Dewhirst, F.E. and Brunetti, A.: Prostaglandin E2 Levels and Human Periodontal Disease, Prostaglandins6:81-85,1974.
19.
Holmes, L.G. and Elattar, T.M.A.: Gingival Inflammation Assessed by Histology, 3H-estrone Metabolism and Prostaglandin E2 Levels, J Periodont Res12:500-509, 1977.
20.
Elattar, T.M.A. : Prostaglandin E2 in Human Gingiva in Health and Disease and Its Stimulation by Female Sex Steroids, Prostaglandins11:331-341, 1976.
21.
Woods, J.F. and Nichols,George, Jr.: Collagenolytic Activity in Rat Bone Cells, J Cell Biod26:747-757, 1965.
22.
Nagai, Y.; Lapiere, C.M.; and Gross, J.: Tadpole Collagenase. Preparation and Purification, Biochem5:3123-3130, 1966,
23.
Narayanan, A.S. and Page, R.C.: Serum Modulates Collagen Types in Human Gingiva Fibroblasts, FEBS Letters80:221-224, 1977.
24.
Barnett, M.L. : The Fine Structure of
25.
Human Connective TissueMast Cells inPeriodontal Disease, J Periodont Res9:84-91, 1974.
26.
Gomes, B.C.; Hausman, E.; Weinfeld, N.; and Deluca, C.: Prostaglandins: Bone Resorption Stimulating Factors Released from Monkey Gingiva , Calcif Tiss Res19 :285-293, 1976.
27.
Das, S.K. ; Adhikary, P.K.; and Bhat-Tacharyya , D.K.: Detection of Prostaglandins in Bovine Gingival Tissue, J Dent Res55:185-188, 1976.
28.
Kafrawy, A.H. , and Mitchell, D.F.: Effects of Prostaglandin E1 on the Periodontium of Rats, J Dent Res56:1132-1133, 1977 .
29.
Blumenkrantz, H.,and Sondergaard , J.: Effect of Prostaglandins E1 and F1α on Biosynthesis of Collagen, Nature (New Biol)239:246,1972.
30.
Raisz, L.G., and Koolemans-Beynen, A.R.: Inhibition of Bone Collagen Synthesis by Prostaglandin E2 in Organ Culture, Prostaglandins8:377-385, 1974.
31.
Kadis, B., and Goodson, J.M.: Hormonal Influences on Bone Cells, J Dent Res56:
32.
A70 (Abstract 110), 1977.
33.
Maganiello, V., and Vaughan, M.: Prostaglandin E1 Effects on Adenosine 3': 5'-cyclic Monophosphate Concentration and Phosphodiesterase Activity in Fibroblasts , Proc Nat Acad Sci69:269-273, 1972.
Bona, C.A.: Fate of Endotoxin in Macrophages : Biological and Ultrastructural Aspects , J Infect Dis128:S74-S81, 1973.
36.
Wahl, L.M.; Wahl, S.M.; Mergenhagen, S.E.; and Martin, G.R.: Collagenase Production by Endotoxin-Activated Macrophages, Proc Nat Acad Sci71:3589-3601, 1974 .
37.
Barbanell, R.; Lian, J.B.; and Keith, D.A.: A Review of Collagen Degradation with Particular Emphasis Upon That Which Occurs During Inflammatory Periodontal Disease, J West Soc Periodontol, Periodontol Abs24: 43-55,1976.
